title: Molecular imaging reveals time course of matrix metalloproteinase activity in acute cutaneous vasculitis in vivo.
abstract: Matrix metalloproteinases (MMPs) play a critical role in various pathological conditions including cutaneous inflammation. Thus far, serial assessment of MMP activity in ongoing inflammation is hampered due to technical limitations. Here, we present an innovative method for longitudinal detection of MMP activity by in vivo imaging. First, we analysed skin sections from patients suffering from leucocytoclastic vasculitis (LcV) and detected a significant MMP signal via immunofluorescence staining. Then, we mimicked LcV in mice in a well-studied model of immune complex-mediated vasculitis (ICV). This acute inflammatory process was serially visualized in vivo using the fluorescence-labelled MMP tracer Cy5.5-AF443. The deposition of fluorescence-labelled immune complexes and MMP tracer distribution was visualized repeatedly and non-invasively by fluorescence reflectance imaging. In correlation with the presence of MMP-2 and MMP-9 in immunofluorescence stainings, Cy5.5-AF443 accumulated in ICV spots in the skin of C57BL/6 mice. This tracer accumulation could also be observed in mice equipped with a dorsal skinfold chamber, where microscopic observations revealed an increased recruitment of fluorescence-labelled leucocytes during ICV. The specificity of the MMP tracer was supported by (i) analysis of mice deficient in functional β2 -integrins (CD18(-/-) ) and (ii) subsequent MMP immunofluorescence staining. These findings let us conclude that MMP accumulation in the acute phase of ICV depends on β2 -mediated leucocyte recruitment. In summary, we show that MMPs are involved in ICV as determined by Cy5.5-AF443, a new optical marker to longitudinally and non-invasively follow MMP activity in acute skin inflammation in vivo.
authors.affiliation: Department of Dermatology, University Hospital of Muenster, Muenster, Germany.
authors.forename: Annika Kathrin
authors.initials: AK
authors.lastname: Steingräber
authors.forename: Sonja
authors.initials: S
authors.lastname: Schelhaas
authors.forename: Andreas
authors.initials: A
authors.lastname: Faust
authors.forename: Andreas Hans
authors.initials: AH
authors.lastname: Jacobs
authors.forename: Michael
authors.initials: M
authors.lastname: Schäfers
authors.forename: Tobias
authors.initials: T
authors.lastname: Goerge
mesh_terms: D000818:Animals; D001183:Arthus Reaction; D018821:CD18 Antigens / Q000378:metabolism; D002232:Carbocyanines / Q000737:chemistry; D005260:Female; D006801:Humans; D007249:Inflammation / Q000378:metabolism; D008297:Male; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008833:Microcirculation; D008856:Microscopy, Fluorescence; D057054:Molecular Imaging / Q000379:methods*; D009504:Neutrophils / Q000201:enzymology*; D012867:Skin / Q000473:pathology; D018366:Vasculitis, Leukocytoclastic, Cutaneous / Q000201:enzymology / Q000473:pathology*
chemical_list: D018821:CD18 Antigens; C098793:CY5.5 cyanine dye; D002232:Carbocyanines; D020782:Matrix Metalloproteinases
keywords: Arthus reaction; intra-vital microscopy; matrix metalloproteinase inhibitor; neutrophils; optical imaging